人组织型激肽释放酶6在卵巢上皮性肿瘤中的表达及其与临床病理特征和预后的关系

Objective To approach the relationship between the expression of hK6 in ovarian neoplasm and clinicopathological variables and prognosis in ovarian cancer patients for finding a new tumor marker of the ovarian cancer. Methods The expression of hK6 was detected by immunohistochemistry in 19 cases of benign, 11 cases of borderline and 45 cases of malignant ovarian neoplasms and statistically analyzed whether its expression correlate with clinicopathological variables and prognosis in patients with ovarian cancer. Results The expression of hK6 in ovarian cancer tissues (60.0%) was significantly higher than that in the benign (15.8%) and borderline (27.3%) ovarian neoplasm tissues (P < 0.01). The expression of hK6 in higher-grade ovarian cancer tissues (68.4%) was higher than that in low-grade ones (14.3%, P<0.05). The expression of hK6 in late-stage (stage Ⅲ ,76.7%) was significantly higher than that in early-stage (stage Ⅰ or Ⅱ ,26.7%, P < 0.01). The expression of hK6 was significantly higher in patients with lymph node metastasis (77.8%) than that in patients without (33.3% ,P<0.01). The expression of hK6 in the cancer tissues in the patients died, or with reeeurence or metastasis within 3 years after surgery was higher (75.0%) than that in the patients with stable disease (42.9%, P < 0. 05). Conclusion The expression of hK6 in ovarian cancer was higher than that in benign and borderline ovarian neoplasms. The expression of hK6 is higher in the ovarian cancer of late stage, higher-grade, with lymph node metastasis and is associated with a poorer prognosis, hK6 may become a new markers in prediction of prognosis of the patients with ovarian tumors.     

荧光实时逆转录PCR定量研究进展

荧光实时逆转录PCR广泛应用于稳态mRNA水平的定量，并且它已经成为基础研究、分子医学和生物技术的一种必不可少的实验工具。用此技术定量具有范围宽、敏感性高、精确度高，无PCR后处理步骤，避免了交叉污染，且产出率高，可以进行多重检测等特点。然而要成功地应用该技术并不是一件容易的事。现对方法学建立、扩增效率以及标准化等方面作一综述。     

外周血IL-18受体的表达对肾移植排斥反应的诊断和治疗的临床意义

目的 检测尿毒症患者肾移植前后外周血白细胞介素18受体(IL-18R)的变化,探讨IL-18R在肾移植急性排斥反应中的作用.方法 受试者分为尿毒症组(n=32),肾功能稳定组(n=24),急性排斥组(n=8),长期存活组(n=12)和健康对照组(n=7).提取受试者的外周血淋巴细胞,加入双荧光标记的鼠抗人CD4/IL-18Rα及CD8/IL-18Rα单克隆抗体,利用流式细胞仪进行测定,计算机分析打印结果.结果 CD4/IL-18Rα及CD8/IL-18Rα在健康对照组(n=7)中的阳性率分别为(5.9±3.4)%和(14.9±4.6)%,而在尿毒症组的阳性率分别为(17.6±9.7)%和(28.0±5.6)%,两组比较差别有显著性(P=0.02,P=0.04).术后恢复顺利者及术后长期存活者外周血CD4/IL-18Rα及CD8/IL-18Rα的表达明显低于急性排斥反应患者.排斥反应发生时外周血CD4/IL-18Rα及CD8/IL-18Rα的阳性率分别为(31.9±11.6)%和(45.7±14.7)%,明显高于肾功能正常组;激素冲击治疗后其阳性率分别为(4.6±1.7)%和(11.2±3.8)%,与激素冲击治疗前比较明显降低.结论 利用流式细胞技术检测尿毒症患者肾移植前后外周血CD4/IL-18Rα及CD8/IL-18Rα的变化,可以较早预测肾移植后急性排斥反应,预测肾移植后恢复情况,评估排斥反应对激素治疗的效果,为排斥反应的早期诊断和选择抗排斥治疗方案提供简单、快速和非创伤性的检测方法.     

SYBR Green实时荧光定量PCR技术平台的建立

目的建立SYBRGreen实时荧光定量PCR技术平台。方法通过摸索SYBRGreen工作液的配制方法,经典PCR检测HBVDNA方法转为SYBRGreen实时荧光定量PCR方法及建立大鼠TGF-bate1mRNA表达水平的SYBRGreen实时荧光定量PCR方法。结果SYBRGreenⅠ工作液,40×H2O溶液稳定性好,最佳反应浓度为1∶10000。HBVDNA的SYBRGreen实时荧光定量PCR方法,最佳的MgCl2反应浓度4.0mmol,引物浓度0.5μmol,81℃时收集荧光信号。定量分析用标准曲线斜率为-3.32,相关系数为-0.994,误差为0.016。大鼠TGF-bate1表达水平的SYBRGreen实时荧光定量PCR方法,TGF-bate1和Bate-actin的最佳MgCl2反应浓度均为3.5mmol,引物浓度前者为0.8μmol,后者为0.5μmol,在87℃时收集荧光信号。靶基因TGF-bate1mRNA的相对量用下述公式计算:(1+Etgf)Ct1/(1+Eact)Ct2。结论初步建成了可检测DNA和mRNA的SYBRGreen实时荧光定量PCR技术平台。     

肾移植患者手术前后外周血CD4＋T淋巴细胞IL-18Rα的表达及意义

目的：探讨外周血CD4^＋T淋巴细胞IL-18Rα表达在肾移植急性排斥反应中的作用及临床意义。方法：采用分别检测肾移植患者手术前、手术后1w及2w外周血CD4^＋T淋巴细胞IL-18Rα表达量，并进行比较分析。结果：肾移植患者手术前外周血CD4^＋T淋巴细胞IL-18Rα表达量明显高于正常对照组（P〈0．01）。急性排斥组手术后1w、2w与手术前比较外周血CD4^＋T淋巴细胞IL-18Rα表达量明显增加（P〈0．01）。稳定组手术后1w与手术前比较外周血CD4^＋T淋巴细胞IL-18Rα表达量明显增加（P〈0．01），手术后2w则下降至手术前水平（P〉0．05）。结论：肾移植患者手术前、手术后存在Th1／Th2失衡，表现为Th1优势应答。肾移植手术前后监测外周血CD4^＋T淋巴细胞IL-18Rα表达量变化对于判断急性排斥反应有一定临床意义。     

PCR基因扩增分析前的质量保证

医学检验的目的是为临床提供可靠的实验诊断依据 ,为了保证检验结果的准确性 ,必须对检测的全过程 (分析前、分析中、分析后 )进行质量控制。在实际工作中对于分析中的质量控制一般比较重视 ,但对分析前的质量控制往往有所忽视 ,例如患者标本留取时机 ,标本收集、处理、储存和运输等是否符合实验要求 ,实验室设置是否合理 ,这些均为影响实验结果的直接因素。文献报告在实验误差中分析前误差约占 70 % ,能否正确规范化收集处理标本是分析前质量保证的重要内容[1 ] ,也是减少实验误差、确保准确及时为临床提供检测结果的重要环节。本文现就临…     

人组织型激肽释放酶6在卵巢上皮性肿瘤中的表达及其与临床病理特征和预后的关系

Objective To approach the relationship between the expression of hK6 in ovarian neoplasm and clinicopathological variables and prognosis in ovarian cancer patients for finding a new tumor marker of the ovarian cancer. Methods The expression of hK6 was detected by immunohistochemistry in 19 cases of benign, 11 cases of borderline and 45 cases of malignant ovarian neoplasms and statistically analyzed whether its expression correlate with clinicopathological variables and prognosis in patients with ovarian cancer. Results The expression of hK6 in ovarian cancer tissues (60.0%) was significantly higher than that in the benign (15.8%) and borderline (27.3%) ovarian neoplasm tissues (P < 0.01). The expression of hK6 in higher-grade ovarian cancer tissues (68.4%) was higher than that in low-grade ones (14.3%, P<0.05). The expression of hK6 in late-stage (stage Ⅲ ,76.7%) was significantly higher than that in early-stage (stage Ⅰ or Ⅱ ,26.7%, P < 0.01). The expression of hK6 was significantly higher in patients with lymph node metastasis (77.8%) than that in patients without (33.3% ,P<0.01). The expression of hK6 in the cancer tissues in the patients died, or with reeeurence or metastasis within 3 years after surgery was higher (75.0%) than that in the patients with stable disease (42.9%, P < 0. 05). Conclusion The expression of hK6 in ovarian cancer was higher than that in benign and borderline ovarian neoplasms. The expression of hK6 is higher in the ovarian cancer of late stage, higher-grade, with lymph node metastasis and is associated with a poorer prognosis, hK6 may become a new markers in prediction of prognosis of the patients with ovarian tumors.     

Tiam1表达与大肠癌外周血微转移的关系

目的〓〖HTK〗探讨Tiam1与大肠癌外周血微转移的关系。〖HTW〗方法〓〖HTK〗采用实时荧光定量逆转录PCR方法检测51例大肠癌患者外周血微转移标志物CK20 mRNA的表达以及Tiam1在大肠癌组织及距肿瘤边缘5～10cm的正常粘膜组织中的表达,应用SPSS11.5统计软件分析Tiam1 mRNA表达水平与大肠癌临床表现及病理特征的相关性。〖HTW〗结果〓〖HTK〗51例中18例（35.29%）检测到外周血微转移。Tiam1 mRNA在有外周血微转移的大肠癌组织及无外周血微转移的大肠癌组织中表达分别为（72.11±2.38）×10-4、（15.78±5.01）×10-4, 差异有统计学意义（P＜0.05）。大肠癌Tiam1 mRNA的表量与患者的性别、年龄、原发部位、肿瘤直径、外周血CEA值无相关关系（P>0.05）, 但与肿瘤外周血微转移、分化程度、浸润深度、淋巴结转移、远处转移、TNM分期相关（P<0.05）。〖HTW〗结论〓〖HTK〗Tiam1与大肠癌外周血微转移、组织分化、淋巴结转移密切相关,检测Tiam1 mRNA含量有助于判断大肠癌的侵袭和转移,尤其是外周血微转移。     

乳腺癌组织KLK7 mRNA的表达及其与临床病理特征的关系

目的建立人KLK7mRNA荧光定量检测方法,对正常乳腺组织和癌组织KLK7 mRNA进行定量,分析其表达与部分临床病理特征的关系,探讨其临床意义。方法应用实时荧光定量PCR检测48例乳腺癌切除的组织中KLK7基因mRNA的表达量,并分析KLK7 mRNA的表达量与年龄、TNM分期、组织学类型、淋巴结转移、雌激素受体（ER）、孕激素受体（PR）等临床病理特征的关系。结果KLK7 mRNA在乳腺正常组织相对表达水平为0．091±0．139,在乳腺癌组织中的相对表达水平为0．107±0．122,无统计学意义（P〉0．05）。但KLK7 mRNA在有淋巴结转移的患者乳腺癌组织中的表达水平高于未转移组（P〈0．05）;在Ⅲ-Ⅳ期患者乳腺癌组织中高于Ⅰ．Ⅱ期患者（P〈0．05）;ER（＋）阳性乳腺癌组织明显低于ER（-）乳腺癌组织（P〈0．05）。KLK7 mRNA表达水平与患者年龄、组织病理分类以及孕激素受体状态无相关性（P〉0．05）。结论乳腺癌组织KLK7 mRNA表达与TNM分期、ER状态及肿瘤转移情况有关。